p53 activated by AND gate genetic circuit under radiation and hypoxia for targeted cancer gene therapy.
p53 activated by AND gate genetic circuit under radiation and hypoxia for targeted cancer gene therapy.
复制标题
辐射和缺氧下与门遗传电路激活p53用于靶向癌症基因治疗
DOI：
10.1111/cas.12739
复制
发表时间：
2015-09
期刊：
影响因子：
5.7
通讯作者：
Wang W
中科院分区：
文献类型：
--
作者：
Ding M;Li R;He R;Wang X;Yi Q;Wang W
放射性基因治疗是近年来发展起来的一种新的肿瘤治疗策略，但治疗基因在肿瘤周围组织中的表达会对正常细胞产生不可接受的毒性。为了限制靶向肿瘤块中的基因表达，我们利用肿瘤细胞的缺氧和辐射耐受特性，通过将辐射敏感性启动子cArG6、热休克反应元件SNF1、HSF1和HSE4与逆转录病毒载体plxsn连接，建立了合成的与门基因电路。通过下游增强型绿色荧光蛋白和wtp53在非小细胞肺癌A549细胞和裸鼠模型中的表达来鉴定它们的构建和动态活性过程。结果表明，较低的辐射剂量（6戈伊）即可激活与门基因电路，激活后的与门基因电路在体内外均能诱导肿瘤细胞产生明显的凋亡效应和生长抑制作用。辐射和缺氧激活的与门基因电路可产生更强的靶向杀瘤活性，为肺腺癌的靶向和有效基因治疗提供了一种新的策略，而低剂量激活的与门基因电路的特性意味着该模型可进一步用于减少临床放射治疗的副作用。
Radio-activated gene therapy has been developed as a novel therapeutic strategy against cancer; however, expression of therapeutic gene in peritumoral tissues will result in unacceptable toxicity to normal cells. To restrict gene expression in targeted tumor mass, we used hypoxia and radiation tolerance features of tumor cells to develop a synthetic AND gate genetic circuit through connecting radiation sensitivity promoter cArG6, heat shock response elements SNF1, HSF1 and HSE4 with retroviral vector plxsn. Their construction and dynamic activity process were identified through downstream enhanced green fluorescent protein and wtp53 expression in non-small cell lung cancer A549 cells and in a nude mice model. The result showed that AND gate genetic circuit could be activated by lower required radiation dose (6 Gy) and after activated, AND gate could induce significant apoptosis effects and growth inhibition of cancer cells in vitro and in vivo. The radiation- and hypoxia-activated AND gate genetic circuit, which could lead to more powerful target tumoricidal activity represented a promising strategy for both targeted and effective gene therapy of human lung adenocarcinoma and low dose activation character of the AND gate genetic circuit implied that this model could be further exploited to decrease side-effects of clinical radiation therapy.